Selected anthranilaminde pyridinamides and their use as pharmaceutical agents

ABSTRACT

Selected anthranilamide pyridinamines of general formula I  
                 
 
     in which R 1  and R 2  have the meanings that are indicated in the description, as VEGFR-2 and VEGFR-3 inhibitors, their production and use as pharmaceutical agents for treating various diseases that are triggered by persistent angiogenesis, are described.

[0001] The invention relates to selected anthranilamide pyridinamines asVEGFR-2 and VEGFR-3 inhibitors, their production and use aspharmaceutical agents for treating diseases that are triggered bypersistent angiogenesis.

[0002] Persistent angiogenesis can be the cause of various diseases,such as psoriasis; arthritis, such as rheumatoid arthritis, hemangioma,angiofibroma; eye diseases, such as diabetic retinopathy, neovascularglaucoma; renal diseases, such as glomerulonephritis, diabeticnephropathy, malignant nephrosclerosis, thrombic microangiopathicsyndrome, transplant rejections and glomerulopathy; fibrotic diseases,such as cirrhosis of the liver, mesangial cell proliferative diseasesand arteriosclerosis, or it can result in an aggravation of thesediseases.

[0003] Persistent angiogenesis is induced by the factor VEGF via itsreceptor. So that VEGF can exert this action, it is necessary that VEGFbind to the receptor, and a tyrosine phosphorylation is induced.

[0004] Direct or indirect inhibition of the VEGF receptor (VEGF=vascularendothelial growth factor) can be used for treating such diseases andother VEGF-induced pathological angiogenesis and vascular permeableconditions, such as tumor vascularization. For example, it is known thatthe growth of tumors can be inhibited by soluble receptors andantibodies against VEGF.

[0005] Anthranilic acid amides that are used as pharmaceutical agentsfor treating psoriasis; arthritis, such as rheumatoid arthritis,hemangioma, angiofibroma; eye diseases, such as diabetic retinopathy,neovascular glaucoma; renal diseases, such as glomerulonephritis,diabetic nephropathy, malignant nephrosclerosis, thrombicmicroangiopathic syndrome, transplant rejections and glomerulopathy;fibrotic diseases, such as cirrhosis of the liver, mesangial cellproliferative diseases, arteriosclerosis, injuries to nerve tissue, andfor inhibiting the reocclusion of vessels after balloon cathetertreatment, in vascular prosthetics or after mechanical devices are usedto keep vessels open, such as, e.g., stents, are known from WO 00/27819.

[0006] The known compounds are generally effective in the indicationscited, but their effectiveness generally accompanies toxicity and aninferior compatibility of the medication.

[0007] There is therefore a desire, on the one hand, for more effectivecompounds, and, on the other hand, for more toxicologically harmlesscompounds, which, moreover, should also be more compatible.

[0008] It has now been found that compounds of general formula I

[0009] in which

[0010] R¹ stands for indazolyl, indolinyl, quinolinyl or for the group

[0011] which optionally can be substituted in one or more places in thesame way or differently with halogen, hydroxy, C₁-C₄-alkyl,C₁-C₄-alkoxy, halo-C₁-C₄-alkyl or with the group ═O or —OR³ or withcyano-C₁-C₃-alkyl,

[0012] R₂ stands for hydrogen or C₁-C₃-alkyl, and

[0013] R³ stands for hydrogen or C₁-C₄-alkyl, with the exception of thecompounds in which R² stands for hydrogen or methyl, and R¹simultaneously stands for unsubstituted indazolyl or quinolinyl, as wellas the enantiomers, racemates, isomers and salts thereof, overcome theabove-indicated drawbacks.

[0014] The compounds according to the invention prevent a tyrosinephosphorylation or stop persistent angiogenesis and thus the growth andpropagation of tumors, whereby they are distinguished in particular by aslighter inhibition of isoforms of Cytochrome P 450 (2C9 and 2C19).

[0015] Pharmaceutical agents that are degraded by these isoforms aregenerally less toxic and better-tolerated.

[0016] Alkyl is defined in each case as a straight-chain or branchedalkyl radical, such as, for example, methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, pentyl, isopentyl, or hexyl, wherebyC₁-C₄-alkyl radicals are preferred.

[0017] Alkoxy is defined in each case as a straight-chain or branchedalkoxy radical, such as, for example, methyloxy, ethyloxy, propyloxy,isopropyloxy, butyloxy, isobutyloxy, or sec-butyloxy.

[0018] Halogen is defined in each case as fluorine, chlorine, bromine oriodine.

[0019] If an acid group is included, the physiologically compatiblesalts of organic and inorganic bases are suitable as salts, such as, forexample, the readily soluble alkali salts and alkaline-earth salts aswell as N-methyl-glucamine, dimethyl-glucamine, ethyl-glucamine, lysine,1,6-hexadiamine, ethanolamine, glucosamine, sarcosine, serinol,tris-hydroxy-methyl-amino-methane, aminopropanediol, Sovak base, and1-amino-2,3,4-butanetriol.

[0020] If a basic group is included, the physiologically compatiblesalts of organic and inorganic acids are suitable, such as hydrochloricacid, sulfuric acid, phosphoric acid, citric acid, tartaric acid,fumaric acid, i.a.

[0021] Those compounds of general formula I, in which

[0022] R¹ stands for indazolyl, indolinyl, quinolinyl or for the group

[0023] which optionally can be substituted in one or more places in thesame way or differently with methyl, methoxy or with the group ═O orwith cyanoethyl, and

[0024] R² stands for hydrogen, with the exception of the compounds inwhich R² stands for hydrogen, and R¹ simultaneously stands forunsubstituted indazolyl or quinolinyl, as well as the enantiomers,racemates, isomers and salts thereof, are especially effective.

[0025] The compounds of general formula I according to the inventionalso contain the possible tautomeric forms and comprise the E- orZ-isomers, or, if a chiral center is present, also the racemates andenantiomers.

[0026] The compounds according to the invention as well as theirphysiologically compatible salts prevent a tyrosine phosphorylation orstop persistent angiogenesis and thus the growth and propagation oftumors, whereby they are distinguished in particular by a slighterinhibition of isoforms of Cytochrome P 450 (2C9 and 2C19). Medicationusing the compounds according to the invention can therefore be done atno risk even without regard to pharmaceutical agents that areadministered at the same time and that are degraded via these isoforms.

[0027] The compounds of formula I as well as their physiologicallycompatible salts can be used as pharmaceutical agents based on theirinhibitory activity relative to the phosphorylation of the VEGFreceptor. Based on their profile of action, the compounds according tothe invention are suitable for treating diseases that are caused orpromoted by persistent angiogenesis.

[0028] Since the compounds of formula I are identified as inhibitors ofthe tyrosine kinases KDR and FLT, they are suitable in particular fortreating those diseases that are caused or promoted by persistentangiogenesis that is triggered via the VEGF receptor or by an increasein vascular permeability.

[0029] The subject of this invention is also the use of the compoundsaccording to the invention as inhibitors of the tyrosine kinases KDR andFLT.

[0030] Subjects of this invention are thus also pharmaceutical agentsfor treating tumors or use thereof.

[0031] The compounds according to the invention can be used either aloneor in a formulation as pharmaceutical agents for treating psoriasis,Kaposi's sarcoma; restenosis, such as, e.g., stent-induced restenosis,endometriosis, Crohn's disease, Hodgkin's disease, leukemia; arthritis,such as rheumatoid arthritis, hemangioma, angiofibroma; eye diseases,such as diabetic retinopathy, neovascular glaucoma; renal diseases, suchas glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis,thrombic microangiopathic syndrome, transplant rejections andglomerulopathy; fibrotic diseases, such as cirrhosis of the liver,mesangial cell proliferative diseases, arteriosclerosis, injuries tonerve tissue, and for inhibiting the reocclusion of vessels afterballoon catheter treatment, in vascular prosthetics or after mechanicaldevices are used to keep vessels open, such as, e.g., stents, asimmunosuppressive agents, for supporting scar-free healing, in senilekeratosis and in contact dermatitis.

[0032] In treating injuries to nerve tissue, quick scar formation on theinjury sites can be prevented with the compounds according to theinvention, i.e., scar formation is prevented from occurring before theaxons reconnect. A reconstruction of the nerve compounds was thusfacilitated.

[0033] The formation of ascites in patients can also be suppressed withthe compounds according to the invention. VEGF-induced edemas can alsobe suppressed.

[0034] Lymphangiogenesis plays an important role in lymphogenicmetastasizing (Karpanen, T. et al., Cancere Res. Mar. 1, 2001, 61(5):1786-90, Veikkola, T., et al., EMBO J. Mar. 15, 2001; 20(6): 1223-31).

[0035] The compounds according to the invention now also show excellentaction as VEGFR kinase 3 inhibitors and are therefore also suitable aseffective inhibitors of lymphangiogenesis.

[0036] By a treatment with the compounds according to the invention, notonly a reduction in the size of metastases but also a reduction in thenumber of metastases is achieved.

[0037] Such pharmaceutical agents, their formulations and uses, are alsosubjects of this invention.

[0038] The invention thus also relates to the use of the compounds ofgeneral formula I for the production of a pharmaceutical agent for useas or for treatment of psoriasis, Kaposi's sarcoma; restenosis, such as,e.g., stent-induced restenosis, endometriosis, Crohn's disease,Hodgkin's disease, leukemia; arthritis, such as rheumatoid arthritis,hemangioma, angiofibroma; eye diseases, such as diabetic retinopathy,neovascular glaucoma; renal diseases, such as glomerulonephritis,diabetic nephropathy, malignant nephrosclerosis, thrombicmicroangiopathic syndrome, transplant rejections and glomerulopathy;fibrotic diseases, such as cirrhosis of the liver, mesangial cellproliferative diseases, arteriosclerosis, injuries to nerve tissue, andfor inhibiting the reocclusion of vessels after balloon cathetertreatment, in vascular prosthetics or after mechanical devices are usedto keep vessels open, such as, e.g., stents, as immunosuppressiveagents, for supporting scar-free healing, in senile keratosis and incontact dermatitis.

[0039] The formation of ascites in patients can also be suppressed withthe compounds according to the invention. VEGF-induced edemas can alsobe suppressed.

[0040] To use the compounds of formula I as pharmaceutical agents, thelatter are brought into the form of a pharmaceutical preparation, whichin addition to the active ingredient for enteral or parenteraladministration contains suitable pharmaceutical, organic or inorganicinert carrier materials, such as, for example, water, gelatin, gumarabic, lactose, starch, magnesium stearate, talc, vegetable oils,polyalkylene glycols, etc. The pharmaceutical preparations can bepresent in solid form, for example as tablets, coated tablets,suppositories, capsules, or in liquid form, for example as solutions,suspensions or emulsions. They also contain, moreover, adjuvants such aspreservatives, stabilizers, wetting agents or emulsifiers, salts forchanging osmotic pressure or buffers.

[0041] For parenteral administration, especially injection solutions orsuspensions, especially aqueous solutions of the active compounds inpolyhydroxyethoxylated castor oil, are suitable.

[0042] As carrier systems, surface-active adjuvants such as salts ofbile acids or animal or plant phospholipids, but also mixtures thereofas well as liposomes or components thereof can also be used.

[0043] For oral administration, especially tablets, coated tablets orcapsules with talc and/or hydrocarbon vehicles or binders, such as forexample, lactose, corn starch or potato starch, are suitable. Theadministration can also be carried out in liquid form, such as, forexample, as juice, to which optionally a sweetener or, if necessary, oneor more flavoring substances, is added.

[0044] The dosage of the active ingredients can vary depending on themethod of administration, age and weight of the patient, type andseverity of the disease to be treated and similar factors. The dailydose is 0.5-1000 mg, preferably 50-200 mg, whereby the dose can be givenas a single dose to be administered once or divided into 2 or more dailydoses.

[0045] The above-described formulations and forms for dispensing arealso subjects of this invention.

[0046] Production of the Compounds According to the Invention

[0047] The following examples explain the production of the compoundsaccording to the invention without the scope of the claimed compoundsbeing limited to these examples.

[0048] The production of the compounds according to the invention iscarried out according to methods that are known in the art, by an amineof general formula II

[0049] in which R¹ has the meaning indicated in general formula I, beingalkylated, and then the amine of general formula II being subjected toreductive alkylation with aldehydes or ketones, whereby it is reacted inthe presence of a reducing agent, such as, for example, sodiumcyanoborohydride, in a suitable inert solvent, such as, for example,ethanol, at temperatures from 0° C. up to the boiling point of thesolvent. An addition of acids such as glacial acetic acid may proveadvantageous.

[0050] If a start is made from a primary amino group, a reaction can beperformed optionally in succession with two different carbonylcompounds, whereby mixed derivatives are obtained [literature, e.g.,Verardo et al. Synthesis (1993), 121; Synthesis (1991), 447; Kawaguchi,Synthesis (1985), 701; Micovic et al. Synthesis (1991), 1043].

[0051] There are also methods to react 1,2-dichloroethane withtriacetoxy borohydride (J. Org. Chem. 1996, 3849). It may beadvantageous first to form the Schiff base by reaction of the aldehydewith the amine in solvents such as ethanol or methanol, optionally withthe addition of adjuvants such as glacial acetic acid, and then to addonly a reducing agent, such as, e.g., sodium cyanoborohydride. TheSchiff base can also be isolated, however, and then reduced to formamine in solvents such as diethyl ether or tetrahydrofuran with reducingagents, such as, for example, lithium aluminum hydride.

[0052] For the production of amides, a start is made from compounds ofgeneral formula

[0053] whereby for this purpose, the processes that are known frompeptide chemistry are available. For example, the corresponding acid inaprotic polar solvents, such as, for example, dimethylformamide, can bereacted via an activated acid derivative that can be obtained, forexample, with hydroxybenzotriazole and a carbodiimide, such as, forexample, diisopropylcarbodiimide, at temperatures of between 0° C. andthe boiling point of the solvent, preferably at 80° C. with the amine.Also, the method of usingO-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (HATU) for the activation provides good yields. As abase, for example, N-methylmorpholine is used, and as a solvent, forexample, dimethylformamide is used. The reaction preferably proceeds atroom temperature. For the amide formation, the process with the mixedacid anhydride, imidazolide or azide can also be used. A priorprotection of an additional amino group, for example as an amide, is notnecessary in all cases, but can advantageously influence the reaction.

[0054] Subjects of this application are thus also compounds of generalformulas II and III

[0055] in which R¹ and R² have the meanings that are indicated ingeneral Formula I, as intermediate products for the production of thecompounds of general formula I.

EXAMPLE 1.0

[0056] Production ofN-(2-Oxo-2,3-dihydro-1H-indol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamide

[0057] 4.5 g (16.9 mmol) of2-amino-N-(2-oxo-2,3-dihydro-1H-indol-6-yl)-benzamide is introduced into90 ml of absolute methanol together with 2.5 g (20.3 mmol) ofpyrid-2-one-5-carboxaldehyde, and it is mixed with 2.5 ml of glacialacetic acid. The feedstock is stirred overnight at room temperature. Itis cooled to 4° C., mixed twice with 750 mg of sodium cyanoborohydrideeach, and stirred for 24 hours. The solid is suctioned off and washedwith methanol and water. It is then absorptively precipitated withsaturated sodium bicarbonate solution and suctioned off again. The solidis dried. 5.16 g (82% of theory) of N-(2-oxo-2,3-dihydro-1H-indol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamideis obtained.

[0058] Similarly produced are also:

Melting Point Example No. R¹ R² [° C.] 1.1

H 1.2

H 1.3

H 1.4

H 1.5

H 1.6

H 1.7

H 1.8

H

EXAMPLE 2.0

[0059] Production ofN-(1-Cyanomethyl-1H-indazol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamide

[0060] 317 mg (1.3 mmol) ofN-(1H-indazol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamideis mixed in 15 ml of dimethylformamide with 280 mg (1.6 mmol) of(6-amino-indazol-1-yl)-acetonitrile, 330 mg of N-methylmorpholine and591 mg (1.55 mmol) ofO-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluoro-phosphate(HATU), and it is stirred for 3 hours at room temperature. The feedstockis then evaporated to the dry state in a vacuum and distributed in 30 mlof ethyl acetate and 30 ml of water, whereby it is also suctioned offfrom the solid. The ethyl acetate phase is dried, filtered andconcentrated by evaporation. The residue is purified on a Flashmastercolumn with a gradient of methylene chloride:ethanol=100:0 to 90:10 asan eluant. 286 mg/55% of theory) ofN-(1-cyanomethyl-1H-indazol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamideis obtained.

EXAMPLE 2.1

[0061] Production ofN-(2-Cyanomethyl-2H-indazol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamide

[0062]N-(2-Cyanomethyl-2H-indazol-6-yl)-2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzamideis produced analogously to Example 2.0.

[0063] Production of Starting Materials

[0064] Process Variant A

[0065] If the production of the intermediate compounds is not described,the latter are known or can be produced analogously to known compoundsor to processes that are described here.

[0066] 2-Methoxypyridine-5-carbaldehyde is a commercially availableproduct (Aldrich)

[0067] ii)

[0068] 2-Pyridone-5carbaldehyde is produced, e.g., according to HoppeSeylers Zeitschrift für physiologische Chemie [Hoppe Seyler's Journalfor Physiological Chemistry], Vol. 325, p. 239, (1961).

[0069] Process Variant B

[0070] 5-Nitro-1,3-dihydro-indol-2-one is produced according to R. T.Courts, J. Org. Chem. 48, 3747, 1970).

[0071] 22.6 g (100 mmol) of 2,4-dinitrophenylacetic acid is dissolved ina mixture of 200 ml of methanol and 830 ml of toluene, mixed at roomtemperature with 83 ml of trimethylsilyldiazomethane (2 molar intoluene; 166 mmol) and stirred for 3 hours at room temperature. Afterconcentration by evaporation to the dry state and drying at 70° C. in avacuum, 24 g (100 % of theory) of 2,4-dinitrophenyl-acetic acid methylester is obtained.

[0072] 26.6 g (83 mmol) of 2,4-dinitrophenylacetic acid methyl ester ishydrogenated in 500 ml of ethanol with 5.4 g of palladium/carbon (10%)under 1 bar of hydrogen for 1.5 hours at room temperature. Aftercatalyst is filtered out, it is concentrated by evaporation. 18.8 g (94%of theory) of 2,4-diaminophenylacetic acid methyl ester is obtained

[0073] 18.8 g (104 mmol) of 2,4-diaminophenylacetic acid methyl ester in500 ml of absolute toluene is mixed at 4° C. with 52 ml of 2-molarsolution of trimethyl aluminum in toluene, and then it is heated for 60minutes to 60° C. After cooling, it is set at pH 7 with 2N sodiumhydroxide solution, filtered on diatomaceous earth and extracted fromethyl acetate. The organic phase is washed, dried, filtered andconcentrated by evaporation. 8.7 g (55% of theory) of6-amino-1,3-dihydro-indol-2-one is obtained.

[0074] 356 mg of 5-nitro-1,3-dihydro-indol-2-one is hydrogenated in 30ml of tetrahydrofuran:ethanol=1:1 with 400 mg palladium on carbon (10%)at room temperature and normal pressure for 1 hour. After catalyst issuctioned off on diatomaceous earth and after concentration byevaporation, 320 mg (100% of theory) of 5-amino-1,3-dihydro-indol-2-oneis obtained.

[0075] In 1 ml of dimethylacetamide, 320 mg of5-amino-1,3-dihydro-indol-2-one is dissolved and mixed drop by drop with371 mg (2 mmol) of 2-nitrobenzoyl chloride, whereby a slight heatingoccurs. After stirring overnight at room temperature, it is concentratedby evaporation in a vacuum, and the residue is taken up in ethyl acetateand water. The suctioning off of an insoluble solid provides 130 mg(21.9% of theory) of2-nitro-N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-benzamide. After shakingout, the organic phase is washed, filtered and concentrated byevaporation, and 400 mg (67% of theory) of2-nitro-N-(2-oxo-2,3-dihydro-1H-indol-5-yl)-benzamide with a meltingpoint of 265° C. is obtained once more.

[0076] Similarly produced is also2-nitro-N-(2-oxo-2,3-dihydro-1H-indol-6-yl)-benzamide with a meltingpoint >300° C.

[0077] Similarly produced to Stage 2 is2-amino-N-(indol-2-on-5-yl)benzoic acid amide with a melting point of219° C.

[0078] Similarly produced to Stage 2 of this example is2-amino-N-(indol-2-on-6-yl)benzoic acid amide with a melting point of230° C.

[0079] Process Variant C

[0080] Production of2-Amino-N-(7-methoxy-2-oxo-2H-chromen-3-yl)-benzamide

[0081] 13 g (85.4 mmol) of 2-hydroxy-4-methoxybenzaldehyde is heated in300 ml of toluene with 9.8 g (102.5 mmol) of n-propylamine hydrochlorideand 11.5 ml (102.5 mmol) of nitroacetic acid ethyl ester for 15 hours ina water separator. 3 ml of nitroacetic acid ethyl ester is then addedagain and boiled for another 5 hours in a water separator. Aftercooling, it is diluted with ethyl acetate and shaken out with water. Theethyl acetate phase is dried, filtered and concentrated by evaporation.The residue is chromatographed on silica gel with methylene chloride asan eluant. 6.14 g (33% of theory) of 3-nitro-7-methoxy-chromen-2-one isobtained.

[0082] In a way similar to Stage 2 from Example B,3-amino-7-methoxy-chromen-2-one is produced from3-nitro-7-methoxy-chromen-2-one in ethanol.

[0083] In a way similar to Stage 3 from Example B,2-nitro-N-(7-methoxybenzopyran-2-on-3-yl)benzoic acid amide is producedfrom 2-nitrobenzoyl chloride and 3-amino-7-methoxy-chromen-2-one [and]2-nitro-N-(7-methoxy-2-oxo-2H-chromen-3-yl)-benzamide.2-Amino-N-(7-methoxy-2-oxo-2H-chromen-3-yl)-benzamide

[0084] In a way similar to Stage 2 of Example B,2-amino-N-(7-methoxy-2-oxo-2H-chromen-3-yl)-benzamide is produced from2-nitro-N-(7-methoxy-2-oxo-2H-chromen-3-yl)-benzamide inethanol:tetrahydrofuran=5:2.

[0085] Similarly produced are also the following intermediate compounds:

Melting Point Example No. R¹ [° C.] i

ii

iii

iv

v

vi

vii

[0086] Process Variant D

[0087] Stage 1

[0088] Analogously to Example1,2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzoic acid methylester is produced from anthranilic acid ester and2-pyridone-5-carbaldehyde in a 72% yield.

[0089] Stage 2

[0090] 450 mg (1.74 mmol) of2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzoic acid methylester is mixed in 20 ml of ethanol with 4 ml of 1N sodium hydroxidesolution, and it is heated for 1 hour to a bath temperature of 120° C.After the ethanol is distilled off in a vacuum, it is diluted with 10 mlof water and set at pH 7 with 2N hydrochloric acid. The precipitatedproduct is suctioned off. 242 mg (57% of theory) of a yield of2-[(6-oxo-1,6-dihydro-pyridin-3-ylmethyl)-amino]-benzoic acid isobtained.

[0091] The sample applications below explain the biological action andthe use of the compounds according to the invention without the latterbeing limited to the examples.

[0092] Solutions Required for the Tests

[0093] Stock solutions

[0094] Stock solution A: 3 mmol of ATP in water, pH 7.0 (−70° C.)

[0095] Stock solution B: g-33P-ATP 1 mCi/100 μl

[0096] Stock solution C: poly-(Glu4Tyr) 10 mg/ml in water

[0097] Solution for dilutions

[0098] Substrate solvent: 10 mmol of DTT, 10 mmol of manganese chloride,100 mmol of magnesium chloride

[0099] Enzyme solution: 120 mmol of tris/HCl, pH 7.5, 10 μM of sodiumvanadium oxide

[0100] Sample Application 1

[0101] Inhibition of the KDR- and FLT-1 Kinase Activity in the Presenceof the Compounds According to the Invention

[0102] In a microtiter plate (without protein binding) that tapers to apoint, 10 μl of substrate mix (10 μl of volume of ATP stock solution A+25 μCi of g-33P-ATP (about 2.5 μl of stock solution B)+30 μl ofpoly-(Glu4Tyr) stock solution C+1.21 ml of substrate solvent), 10 μl ofinhibitor solution (substances corresponding to the dilutions, 3% DMSOin substrate solvent as a control) and 10 μl of enzyme solution (11.25μg of enzyme stock solution (KDR or FLT-1 kinase) are added at 4° C. in1.25 ml of enzyme solution (dilute). It is thoroughly mixed andincubated for 10 minutes at room temperature. Then, 10 μl of stopsolution (250 mmol of EDTA, pH 7.0) is added, mixed, and 10 μl of thesolution is transferred to a P 81 phosphocellulose filter. Then, it iswashed several times in 0.1 M phosphoric acid. The filter paper isdried, coated with Meltilex and measured in a microbeta counter.

[0103] The IC50 values are determined from the-inhibitor concentration,which is necessary to inhibit the phosphate incorporation to 50% of theuninhibited incorporation after removal of the blank reading(EDTA-stopped reaction).

[0104] The results of the kinase inhibition IC50 in μM are presented inthe table below: VEGFR II (KDR) Example No. [μM] 1.0 0.05

[0105] Sample Application 2

[0106] Cytochrome P450 Inhibition

[0107] The Cytochrome P450 inhibition was performed according to thepublication of Crespi et al. (Anal. Biochem., 248, 188-190 (1997)) withuse of baculovirus/insect cell-expressed, human Cytochrome P 450isoenzymes (1A2, 2C9, 2C19, 2D6, 3A4).

[0108] The results are presented in the following table. Inhibition ofthe Cytochrome P450 Isoenzymes (IC50, μM) Cytochrome P450 Isoenzyme 1A22C9 2C19 2D6 3A4 Example 5.2 0.2 0.05 >30 3.6 2.54 of WO 00/27819Example 1.0 >30 23 >30 >30 >30

[0109] The superior action of the compounds according to the inventioncompared to the known compounds can be seen clearly from the result.

[0110] Without further elaboration, it is believed that one skilled inthe art can, using the preceding description, utilize the presentinvention to its fullest extent. The preceding preferred specificembodiments are, therefore, to be construed as merely illustrative, andnot limitative of the remainder of the disclosure in any way whatsoever.

[0111] In the foregoing and in the examples, all temperatures are setforth uncorrected in degrees Celsius and, all parts and percentages areby weight, unless otherwise indicated.

[0112] The entire disclosures of all applications, patents andpublications, cited herein and of corresponding German application No.102 28 090.8, filed Jun. 19, 2002, and U.S. Provisional ApplicationSerial No. 60/404,773, filed Aug. 21, 2002, are incorporated byreference herein.

[0113] The preceding examples can be repeated with similar success bysubstituting the generically or specifically described reactants

1. Compounds of general formula I

in which R¹ stands for indazolyl, indolinyl, quinolinyl or for the group

which optionally can be substituted in one or more places in the sameway or differently with halogen, hydroxy, C₁-C₄-alkyl, C₁-C₄-alkoxy,halo-C₁-C₄-alkyl or with the group ═O or —OR³ or with cyano-C₁-C₃-alkyl,R² stands for hydrogen or C₁-C₃-alkyl, and R³ stands for hydrogen orC₁-C₄-alkyl, with the exception of the compounds in which R² stands forhydrogen or methyl, and R¹ simultaneously stands for unsubstitutedindazolyl or quinolinyl, as well as the enantiomers, racemates, isomersand salts thereof:
 2. Compounds of general formula I, in which R¹ standsfor indazolyl, indolinyl, quinolinyl or for the group

which optionally can be substituted in one or more places in the sameway or differently with methyl, methoxy or with the group ═O or withcyanoethyl, and R² stands for hydrogen, with the exception of thecompounds in which R stands for hydrogen, and R¹ simultaneously standsfor unsubstituted indazolyl or quinolinyl, as well as the enantiomers,racemates, isomers and salts thereof.
 3. Pharmaceutical agents thatcontain at least one compound according to claims 1 and
 2. 4.Pharmaceutical agents according to claim 3 for treating tumors,psoriasis, Kaposi's sarcoma, restenosis, stent-induced restenosis,endometriosis, Crohn's disease, Hodgkin's disease, leukemia, arthritis,rheumatoid arthritis, hemangioma, angiofibroma, eye diseases, diabeticretinopathy, neovascular glaucoma, renal diseases, glomerulonephritis,diabetic nephropathy, malignant nephrosclerosis, thrombicmicroangiopathic syndrome, transplant rejections, glomerulopathy,fibrotic diseases, cirrhosis of the liver, mesangial cell proliferativediseases, arteriosclerosis, injuries to nerve tissue, and for inhibitingthe reocclusion of vessels after balloon catheter treatment, in vascularprosthetics or after the use of stents to keep vessels open, and ofimmune diseases as immunosuppressive agents, and for supportingscar-free healing in senile keratosis and in contact dermatitis. 5.Compounds according to claims 1 and 2 and pharmaceutical agents,according to claims 3 and 4, with suitable formulations and vehicles. 6.Use of the compounds of formula I, according to claims 1 and 2, asinhibitors of the tyrosine kinases KDR and FLT.
 7. Use of the compoundsof general formula I, according to claims 1 and 2, in the form of apharmaceutical preparation for enteral, parenteral and oraladministration.
 8. Use of the compounds of general formula I, accordingto claims 1 and 2, as VEGFR-kinase 3 inhibitors in lymphangiogenesis. 9.Use of the compounds of general formula I, according to claims 1 and 2,for suppressing the formation of ascites and for suppressingVEGF-induced edemas.
 10. Compounds of general formulas II and III

in which R¹ and R², which have the meanings indicated in general formulaI, as intermediate products for the production of compounds of generalformula I.